AML1/ETO recruits DNMTs, HDAC1, and MeCP2 to the RARβ2 promoter. (A) ChIPs were performed in mock, A/E-HA, and SKNO-1 cells using the indicated antibodies or in absence of antibody (no-Ab) or the anti-ETO antibody (B) and analyzed by PCR with primer for the βRARE binding site on RARβ2 promoter or for the AML1 binding site on p14^ARF promoter. A region including the p16^INK4a transcriptional start site and RARβ2 exon 7 (EX7), each lacking the AML1 and the RARE binding sites, respectively, were amplified to evaluate the specificity of AML1/ETO binding. (C-D) ChIPs were performed on mock, A/E-HA, and SKNO-1 cell lines using antibodies specific for MeCP2, DNMT1, DNMT3a, DNMT3b, and HDAC1 or with no antibody (no-Ab) as a negative control. Immunoprecipitated chromatins were analyzed by PCR using primer pairs specific for the amplification of the promoter region containing the βRARE and the 5-UTR exon 1 sequences on RARβ2 gene (RARβ2pr) or regions surrounding the AML1 site of the interleukin-3 promoter (IL-3pr). SKNO-1 cells were treated or not with 1 μM RA for 24 hours. Samples representing 0.02% of sonicated chromatin (input) were included in the PCR analysis. The amplification of a GAPDH coding region was used as a control of nonspecific precipitated sequences.